CN107418910A - A kind of biological agent for preventing and treating crop in cruciferae clubroot and its application - Google Patents
A kind of biological agent for preventing and treating crop in cruciferae clubroot and its application Download PDFInfo
- Publication number
- CN107418910A CN107418910A CN201710312344.XA CN201710312344A CN107418910A CN 107418910 A CN107418910 A CN 107418910A CN 201710312344 A CN201710312344 A CN 201710312344A CN 107418910 A CN107418910 A CN 107418910A
- Authority
- CN
- China
- Prior art keywords
- bacillus amyloliquefaciens
- cruciferae
- crop
- biological agent
- clubroot
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000003124 biologic agent Substances 0.000 title claims abstract description 62
- 241000219193 Brassicaceae Species 0.000 title claims abstract description 60
- 241000894006 Bacteria Species 0.000 claims abstract description 43
- 239000002689 soil Substances 0.000 claims abstract description 32
- 241000196324 Embryophyta Species 0.000 claims abstract description 27
- 238000000034 method Methods 0.000 claims abstract description 23
- 238000010438 heat treatment Methods 0.000 claims abstract description 16
- 238000004519 manufacturing process Methods 0.000 claims abstract description 15
- 230000001580 bacterial effect Effects 0.000 claims abstract description 13
- 238000003973 irrigation Methods 0.000 claims abstract description 12
- 230000002262 irrigation Effects 0.000 claims abstract description 12
- 238000002360 preparation method Methods 0.000 claims description 43
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 20
- 235000013311 vegetables Nutrition 0.000 claims description 6
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 4
- 230000000717 retained effect Effects 0.000 claims description 4
- 241000726221 Gemma Species 0.000 claims description 3
- 238000009331 sowing Methods 0.000 claims description 2
- 241000193744 Bacillus amyloliquefaciens Species 0.000 abstract description 24
- 230000002265 prevention Effects 0.000 abstract description 19
- 238000010790 dilution Methods 0.000 description 29
- 239000012895 dilution Substances 0.000 description 29
- 244000221633 Brassica rapa subsp chinensis Species 0.000 description 20
- 235000010149 Brassica rapa subsp chinensis Nutrition 0.000 description 20
- 230000000694 effects Effects 0.000 description 19
- 239000007788 liquid Substances 0.000 description 18
- 241000219198 Brassica Species 0.000 description 15
- 238000000855 fermentation Methods 0.000 description 15
- 230000004151 fermentation Effects 0.000 description 15
- 201000010099 disease Diseases 0.000 description 13
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 13
- 235000003351 Brassica cretica Nutrition 0.000 description 12
- 235000003343 Brassica rupestris Nutrition 0.000 description 12
- QKSKPIVNLNLAAV-UHFFFAOYSA-N bis(2-chloroethyl) sulfide Chemical compound ClCCSCCCl QKSKPIVNLNLAAV-UHFFFAOYSA-N 0.000 description 12
- 239000003085 diluting agent Substances 0.000 description 12
- 235000010460 mustard Nutrition 0.000 description 12
- 238000012545 processing Methods 0.000 description 12
- 238000002474 experimental method Methods 0.000 description 11
- 239000000243 solution Substances 0.000 description 11
- 240000007124 Brassica oleracea Species 0.000 description 10
- 235000003899 Brassica oleracea var acephala Nutrition 0.000 description 10
- 235000011301 Brassica oleracea var capitata Nutrition 0.000 description 10
- 235000001169 Brassica oleracea var oleracea Nutrition 0.000 description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 8
- 230000000443 biocontrol Effects 0.000 description 8
- 244000005700 microbiome Species 0.000 description 8
- 238000009472 formulation Methods 0.000 description 7
- 238000003306 harvesting Methods 0.000 description 7
- 239000000203 mixture Substances 0.000 description 7
- 238000011835 investigation Methods 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 238000010998 test method Methods 0.000 description 5
- 241001503436 Plasmodiophora brassicae Species 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 235000011331 Brassica Nutrition 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 241000607479 Yersinia pestis Species 0.000 description 3
- 229940041514 candida albicans extract Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 230000003628 erosive effect Effects 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 239000000575 pesticide Substances 0.000 description 3
- 239000004033 plastic Substances 0.000 description 3
- 229920003023 plastic Polymers 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000012138 yeast extract Substances 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- 229920000936 Agarose Polymers 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 208000035240 Disease Resistance Diseases 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 244000038559 crop plants Species 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 239000012137 tryptone Substances 0.000 description 2
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 1
- 241001019659 Acremonium <Plectosphaerellaceae> Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000000536 Brassica rapa subsp pekinensis Nutrition 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 241001655322 Streptomycetales Species 0.000 description 1
- 108010006785 Taq Polymerase Proteins 0.000 description 1
- 241000223259 Trichoderma Species 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000000980 acid dye Substances 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 231100001261 hazardous Toxicity 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 244000000042 obligate parasite Species 0.000 description 1
- 238000000879 optical micrograph Methods 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 230000010496 root system development Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Environmental Sciences (AREA)
- Toxicology (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of biological agent for preventing and treating crop in cruciferae clubroot and its application, belong to microbe field.The production bacterial strain of biological agent is Bacillus amyloliquefaciens strainBacillus amyloliquefaciens KM 68, has been deposited in China typical culture collection center, and deposit number is CCTCC NO:M 2017228.The inventive method is that the heat treatment of the soil sun combines Bacillus amyloliquefaciens strainBacillus amyloliquefaciens KM 68 bacterium solution root irrigation method, this method can effectively preventing crop in cruciferae clubroot, be advantageous to crop in cruciferae volume increase, and the prevention and controls are simple to operate, and expense is low, and biological agent is easy to plant produced.
Description
Technical field
The present invention relates to a kind of biological agent for preventing and treating crop in cruciferae clubroot and its application, belong to microbe
Field.
Background technology
Crop in cruciferae vegetables clubroot is by Brassica genus plasmodiophora brassicae（PLasmodiophora brassicae
Woron）A kind of worldwide soil-borne disease caused by dip-dye.There is generation current most of countries and regions.Due to the pathogen
Infectiousness is strong, and spread speed is fast, and preventing and treating is difficult, and in recent years, crop in cruciferae clubroot is climing rapidly in China's most area
Prolong, serious threat the production of Cruciferae crops.
At present, preventing and treating the method for crop in cruciferae clubroot mainly includes：Chemical control, breeding for disease resistance, biological control.
Because Brassica genus plasmodiophora brassicae belongs to obligate parasite, breeding for disease resistance is that preventing and treating crop in cruciferae clubroot is maximally effective in theory
Mode, however, Brassica genus plasmodiophora brassicae biological strain is numerous, variation is fast, it is difficult to cultivate permanently effective disease-resistant variety, currentization
It is still to prevent and treat the main path of crop in cruciferae clubroot to learn chemical control, but chemical control is not only wasted time and energy, and long
Phase uses chemical pesticide, certainly will cause a large amount of accumulation of chemical pesticide residual in soil, and chemical pesticide residual once enters agricultural product
In, it will directly endanger people health.There is the preventing and treating that biocontrol microorganisms are applied to crop in cruciferae clubroot by researcher in recent years
In, and it is found that some bacterium, such as fungies, bacillus amyloliquefaciens for having good result to prevent and treat to clubroot, bacillus subtilis
Bacterium, streptomycete, Trichoderma, Acremonium endogenetic fungus etc..But biocontrol microorganisms preventing and treating clubroot is used alone, often prevention effect is not
It is good, it is impossible to improve the yield of crops, and long-term use biocontrol microorganisms, the microbial diversity in soil certainly will be caused to change
Become, edaphon is exerted an adverse impact.
The content of the invention
In view of the deficiencies of the prior art, the present invention provides it is a kind of prevent and treat crop in cruciferae clubroot biological agent,
Can efficient, nontoxic, safety, noresidue, it is environment amenable preventing and treating Cruciferae clubroot.
The microbial strains that the present invention uses is bacillus amyloliquefaciensBacillus amyloliquefaciens KM
68 bacterial strains, the bacterial strain are deposited in China typical culture collection center, preservation address on May 2nd, 2017：Wuhan, China,
Wuhan University, deposit number are CCTCC NO: M 2017228.
The bacillus amyloliquefaciens of the present inventionBacillus amyloliquefaciens The bacterial strains of KM 68, from Yunnan Province elder brother
Obtained in the soil of Ming Shi Songming Counties using dilution-plate method and plate streak separation, it is automatic through morphology, cultural colony and DNA
Identification systems determine, and the bacterial strain is a new strains of bacillus amyloliquefaciensBacillus amyloliquefaciens KM
68。
The present invention is another object is that by bacillus amyloliquefaciensBacillus amyloliquefaciensThe bacterial strains of KM 68
Biological agent is made to apply in crop in cruciferae clubroot is prevented and treated.
The application process of the biological agent of described preventing and treating crop in cruciferae clubroot, is comprised the following steps that：
（1）The soil sun is heat-treated：Deep ploughing soil, pour water, seal, then carry out the sun and be heat-treated more than 30 days；
（2）Greenhouse production：Acquisition step（1）The pedotheque of gained sun heat treatment is placed in the hole tray in greenhouse, in hole tray
The vegetable seeds of middle sowing crop in cruciferae, the pedotheque covering for taking the sun to be heat-treated, watering, the plant of crop in cruciferae
Thing seed sprouting cuts off unnecessary seedling to when growing 3 ~ 4 leaves, and one plant of healthy seedling is retained per cave, is made using biology
Agent carries out root irrigation 1 time to crop in cruciferae plant seedlings；
The biological agent is bacillus amyloliquefaciensBacillus amyloliquefaciens The bacterium solution of the bacterial strains of KM 68, bacterium
The gemma amount of liquid is 4 × 105~ 1×107cfu/mL；
The biological agent of preventing and treating crop in cruciferae clubroot provided by the invention has the following advantages that：
（1）The present invention uses soil sun heat treating process combination bacillus amyloliquefaciensBacillus amyloliquefaciens
KM 68 biological agent facture, crop in cruciferae clubroot not only can be effectively prevented and treated, and the production of crop can be significantly improved
Amount；
（2）Present invention application concentration of bacterium solution in biological agent processing procedure is low, and bacterium solution application dosage is few, and only needs pouring root
Once；Simple to operate, expense is low, can have necessarily to the soil environment of deterioration to environment non-hazardous, and by sun heat treatment
Repair.
Brief description of the drawings
Fig. 1 is bacillus amyloliquefaciens strain of the present inventionBacillus amyloliquefaciensKM 68 bacterium colony figure；
Fig. 2 is bacillus amyloliquefaciens strain of the present inventionBacillus amyloliquefaciensKM 68 electron microscope is swept
Trace designs piece；
Fig. 3 is bacillus amyloliquefaciens strain of the present inventionBacillus amyloliquefaciensKM 68 growth curve.
Embodiment
Below in conjunction with drawings and examples to the further detailed description of the present invention, but protection scope of the present invention is not
It is only limitted to the content.
Bacillus amyloliquefaciens strainBacillus amyloliquefaciensKM 68 is from Yunnan Province Songming County soil
It is middle to be obtained using dilution-plate method and plate streak separation screening, Chinese Typical Representative culture guarantor is stored on May 2nd, 2017
Tibetan center, deposit number are CCTCC NO: M 2017228.
Embodiment 1：Bacillus amyloliquefaciens strainBacillus amyloliquefaciensKM 68 screening separation
LB culture medium prescriptions：The g/L of tryptone 10, the g/L of yeast extract 5, the g/L of sodium chloride 5, the g/L of agarose 15；PH values
For 7.4 ± 0.1；Add distilled water stirring and dissolving after weighing, be distributed into triangular flask, in 121 DEG C of autoclave sterilizations, be down flat plate；
The pedotheque of 1 g Yunnan Province Songming County collection is weighed, is put into the triangular flask with bead for filling 50 mL sterilized waters
In, concussion shake up 30 min, 1 mL supernatants are then drawn from triangular flask using liquid-transfering gun, add another mL of Zhi Shengyou 9 without
In the test tube of bacterium water, mixing, according to said method, diluted concentration is respectively prepared as the 10 of original solution concentration-3、10-4、10-5Soil again
Solution, take 100 uL dilute the soil liquid be coated on LB culture medium flat plates, each dilution factor is repeated 3 times, 37 DEG C, 150
The h of shaking table culture 10 under the conditions of r/min, picking single bacterium are fallen within and rule on LB culture mediums, and colony growth situation is observed in timing, then
Using plate streak, bacterial strain is purified, makes inclined-plane label respectively, is stored in the refrigerator that temperature is 4 DEG C.Pass through molecular system
Auxology is analyzed, and is named as bacillus amyloliquefaciens strainBacillus amyloliquefaciensKM 68, deliver to " Chinese allusion quotation
Type culture collection " (abbreviation CCTCC) preserves.
Embodiment 2：Bacillus amyloliquefaciens strainBacillus amyloliquefaciensKM 68 morphology
Bacillus amyloliquefaciens strain is prepared using gradient dilution methodBacillus amyloliquefaciensKM 68 bacterium is hanged
Liquid, the bacteria suspension for drawing 10 uL difference dilution factors respectively is coated on LB culture medium flat plates and cultivated, to growing after 8 ~ 15 h
Single bacterium colony size, shape, surface, edge, the characteristic such as color and transparency observed；
Cultivate the bacillus amyloliquefaciens strain of different dilution factors respectively on LB culture mediumsBacillus amyloliquefaciensKM 68, after cultivating 8 ~ 15 hours, 10uL bacteria suspensions is drawn respectively and carry out film-making, in optical microphotograph
Microscopic observation bacillus amyloliquefaciens strainBacillus amyloliquefaciensKM 68 growing state and morphological feature；
There is following feature by cultivating and observing thalline：Bacterium colony is rounded or oval, has fold, slightly swells, opaque, is in
White, Gram's staining are positive, and are in rod-short under microscope（Bacterium colony figure is shown in Fig. 1, and electron microscope scanning picture is shown in figure
2）.
Embodiment 3：Bacillus amyloliquefaciens strainBacillus amyloliquefaciensKM 68 16s rDNA bases
Because of Sequence Identification
Single bacterium colony is inoculated in LB medium cultures after 24 h, takes 1 uL to carry out bacterium colony PCR checkings.For bacterium solution PCR's
Primer is as follows：Forward primer 27F： 5,-AGAGTTTGATCMTGGCTCAG-3,；Reverse primer 1492R： 5,-
GGTTACCTTGTTACGACTT-3,.PCR reaction systems：The uL of 10xTaq Buffer 2.5, the uL of forward primer 0.5, reversely draw
The uL of thing 0.5, bacterium solution 1 uL, dNTP（10 mmol/L ）The U of 0.5 uL, Taq DNA polymerase 0.5, add water to 25
UL, PCR amplification condition:According to 94 DEG C, 30 s, 57 DEG C, 120 s, 72 DEG C, 90 s program progress after 94 DEG C of min of pre-degeneration 3
35 circulations, last 72 DEG C re-extend 10 min.Electrophoresis uses agarose gel electrophoresis, nucleic acid dye EB, DNA Mark length
Spend for 2000, the PCR products purified finally are delivered into Yunnan Shuo Qing bio tech ltd is sequenced, by acquisition
Strain KM 68 16S rDNA sequences are contrasted with existing correlated series in NCBI nucleic acid databases.As a result show
The strain KM 68 of acquisition withBacillus amyloliquefaciensStrain DM-54,Bacillus amyloliquefaciensStrain Lx-11,Bacillus amyloliquefaciensStrain LD5,Bacillus amyloliquefaciensStrain EB19 similarities reach 99%（It is shown in Table 1）, it may be determined that strain KM 68 belong to
A kind of new strains of bacillus, are named as bacillus amyloliquefaciens strainBacillus amyloliquefaciens KM
68,
。
Embodiment 4：Bacillus amyloliquefaciens strainBacillus amyloliquefaciensKM 68 culture growth
LB culture medium prescriptions are：The g of peptone 10, the g of sodium chloride 5, the g of yeast extract 10, the L of distilled water 1, pH value 7.4；
Use LB Liquid Culture based assays bacillus amyloliquefaciensBacillus amyloliquefaciens KM 68 grows song
Line：The g of tryptone 1 is weighed, the g of yeast extract 0.5, the g of sodium chloride 0.5, agarose 1.5, is dissolved in water and is settled to 100 mL；
Using 11mol/L sodium hydroxide solution by 7.0 ± 0.1, then by the LB culture mediums of configuration, it is distributed into 10 test tubes,
Autoclave sterilization under the conditions of 121 DEG C, room temperature is then cooled to, in an aseptic environment, 10uL Xie Dian is accessed in each test tube
AfnyloliquefaciensBacillus amyloliquefaciens The bacterium solutions of KM 68, under the conditions of 37 DEG C, 150 r/min, shaking table training
Support 24 hours, the every 2 hours OD values for surveying a bacterium solution, with the time（Hour）For abscissa, using OD values as ordinate, solution starch is done
BacillusBacillus amyloliquefaciens The growth curve charts of KM 68（See Fig. 3）, the results showed that, solve starch gemma
BacillusBacillus amyloliquefaciens KM 68 is most fast in 0-10 hours reproduction speed, and subsequent bacterium colony tends towards stability；
It is calculated in mass percent, liquid fermentation medium formula：Analysis for soybean powder 3%, corn flour 3%, sodium chloride 0.5%, calcium carbonate
0.5%, remaining is water；
By bacillus amyloliquefaciens strainBacillus amyloliquefaciensKM 68 spore is transplanted in LB culture mediums,
10 h are cultivated under the conditions of being 37 DEG C in temperature, obtain seed liquor, are 1 according to seed liquor and the volume ratio of water:100 ratio, will
Seed liquor is inoculated into liquid fermentation medium, and 10 h are cultivated under the conditions of being 37 DEG C in cultivation temperature, after the completion of culture, fermentation
Bacillus amyloliquefaciens bacterial concentration is reachable in liquid（1.6~3.2）×108cfu/mL；
Liquid fermentation preparation（Prevent and treat the biological agent of crop in cruciferae clubroot）：Bacteria containing amount is determined through plate count, containing bacterium
Amount reaches（1.6~3.2）×108 cfu/mL。
Embodiment 5：Liquid fermentation preparation（Prevent and treat the biological agent of crop in cruciferae clubroot）To crop in cruciferae
（Pakchoi）The prevention effect experiment and volume increase experiment of clubroot
Experimental subjects：Pakchoi
Prepare biological agent：Experiment biological agent is prepared according to the culture growing method of embodiment 4, is contained through plate count measure
Bacterium amount, the bacteria containing amount of bacillus amyloliquefaciens reach（1.6~3.2）×108cfu/mL；
By biological agent dilution, 20 times obtain preparation A, and by biological agent dilution, 100 times obtain preparation B, and biological agent is diluted
500 times obtain formulation C；
For medicine preparation and processing：
（1）The soil sun is heat-treated：Pour water the soil of deep ploughing 30min, makes the abundant saturation of the moisture in soil, covers one layer of modeling
Material film is sealed, and is then carried out the sun and is heat-treated 30 days, collection sun heat treatment pedotheque；
（2）Greenhouse production：Hole tray is cultivated using 72 holes, by step（1）The cave that the pedotheque of sun heat treatment is placed in greenhouse
In disk, 3 ~ 4 crop in cruciferae are sowed in every cave of hole tray（Pakchoi）Vegetable seeds, take step（1）Solar heat at
Manage pedotheque covering, watering, crop in cruciferae plant（Pakchoi）For seed sprouting to 3 ~ 4 leaves are grown, it is unnecessary to cut off
Seedling, per cave retain one plant of healthy seedling, using the dilution of biological agent to crop in cruciferae（Pakchoi）Seedling
Carry out root irrigation 1 time, the dilution for the biological agent that each plant root adds is 10mL；
A：The dilution of biological agent is the preparation A of biological 20 times of preparation diluent；
B：The dilution of biological agent is the preparation B of biological 100 times of preparation diluent；
C：The dilution of biological agent is the formulation C of biological 500 times of preparation diluent；
D：Blank control, using clear water to crop in cruciferae plant（Pakchoi）Seedling carries out root irrigation 1 time, each plant
The clear water amount that root adds is 10mL；
Test method：Experiment sets 4 processing altogether, 3 repetitions of each processing, Gong12Ge areas, is independent of each other between each area, during harvest
It is each to distinguish survey production；
Investigation method：Investigation each handles totally 108 plants of seedling in 3 areas, treats that to 40 days, seedling is taken out for growth of seedling, surveys children
The overground part fresh weight of seedling；
Clubroot is investigated using the size of root tumour as foundation, with reference to army of department（2009）Identify the morbidity of each plant clubroot
Grade：
0 grade：Root is normal without tumour, root system development；
1 grade：Main root is not fallen ill or unobvious, diameter≤2 times basal part of stem；
2 grades：The obvious enlargement of main root.Diameter≤2-3 times of stem foot；
3 grades：The obvious enlargement of main root, diameter≤3- times of stem foot, no fibrous root；
The total strain number of the incidence of disease=disease plant/statistics
Disease index=∑（Diseased plant numbers at different levels × this plant of disease index）/ (investigating total strain tree × superlative degree value)
Relative control effect=（Compare disease index-processing disease index）× 100/ control disease index
Experimental result is as shown in table 1.
Comparative example 1：The only prevention and controls of soil sun heat treatment
Experimental subjects：Pakchoi
（1）The soil sun is heat-treated：Deep ploughing soil, pour water 30min, makes the abundant saturation of the moisture in soil, covers one layer of plastics
Film is sealed, and is then carried out the sun and is heat-treated 30 days, the pedotheque of collection sun heat treatment；
（2）Greenhouse production：Hole tray is cultivated using 72 holes, by step（1）The cave that is placed in greenhouse of sun heat treatment pedotheque
In disk, 3 ~ 4 crop in cruciferae plants are sowed in every cave of hole tray（Pakchoi）Seed, take step（1）Solar heat at
Manage pedotheque covering, watering, crop in cruciferae plant（Pakchoi）For seed sprouting to 3 ~ 4 leaves are grown, it is unnecessary to cut off
Seedling, per cave retain one plant of healthy seedling；
Test method：Field management produces with conventional；Experiment sets 1 processing, 3 repetitions, totally 3 areas, each distinguishes during harvest
Open survey production；
Investigation method is same as Example 5；Experimental result is as shown in table 1.
Comparative example 2：Only with the prevention and controls of liquid fermentation preparation
Experimental subjects：Pakchoi
Using 72 holes culture hole tray, pedotheque is added in the hole tray in greenhouse, 3 ~ 4 cruciate flowers are sowed in every cave of hole tray
Section's crop plants（Pakchoi）Seed, take pedotheque to cover, water, crop in cruciferae plant（Pakchoi）Seed sprouting is extremely
3 ~ 4 leaves are grown, cut off unnecessary seedling, one plant of healthy seedling is retained per cave；Using the dilution of biological agent to ten
Zi Hua sections crop plants（Pakchoi）Seedling carries out root irrigation 1 time, the dilution for the biological agent that each plant root adds
For 10mL；
E：The dilution of biological agent is the preparation A of biological 20 times of preparation diluent；
F：The dilution of biological agent is the preparation B of biological 100 times of preparation diluent；
G：The dilution of biological agent is the formulation C of biological 500 times of preparation diluent；
Test method：In addition to dispenser sets varying level, other field management produce with conventional；Test and set 3 processing, totally 9
Area, survey production is often distinguished during harvest；
Investigation method is same as Example 5；Experimental result is as shown in table 1；
Experimental result：
Prevention effect of the table 1. to pakchoi clubroot
As a result show：The yield of pakchoi seedling is individually significantly improved using sun heat treatment soil（P<0.05）, but to root
The prevention effect for the disease that swells is only 46.9%, and liquid fermentation preparation is used alone（Prevent and treat the biology system of crop in cruciferae clubroot
Agent）Also the generation of clubroot can be suppressed, but liquid fermentation preparation is used alone（Prevent and treat the biology system of crop in cruciferae clubroot
Agent）Chinese cabbage yield can not be increased（P>0.05）, and the liquid fermentation preparation of various concentrations（Prevent and treat crop in cruciferae clubroot
Biological agent）There is notable difference to the prevention effect of clubroot, when liquid fermentation preparation（Prevent and treat crop in cruciferae clubroot
Biological agent）Middle bacillus amyloliquefaciensBacillus amyloliquefaciensKM 68 bacteria containing amount is respectively 4 ×
105 cfu/mL、2×106 cfu/mL、107Cfu/mL, the prevention effect to clubroot is respectively 35.3%, 64.8%,
77.13%, the yield of sun heat treatment soil combination biocontrol microorganisms, not only obvious increase pakchoi（P<0.05）, and in liquid fermentation
Preparation（Prevent and treat the biological agent of crop in cruciferae clubroot）Bacillus amyloliquefaciensBacillus amyloliquefaciensAlso preventing and treating effect well can be reached when KM 68 bacteria containing amount is relatively low to the clubroot of pakchoi
Fruit, work as bacillus amyloliquefaciensBacillus amyloliquefaciensKM 68 bacteria containing amount reaches 2 × 106cfu/mL
When, 81.7% is reached to the prevention effect of pakchoi clubroot, continues to improve biological and ecological methods to prevent plant disease, pests, and erosion bacteria concentration, the preventing and treating to pakchoi clubroot
Effect improves and unobvious.
Embodiment 6：Liquid fermentation preparation（Prevent and treat the biological agent of crop in cruciferae clubroot）To crop in cruciferae
（Stem mustard）The preventing and treating experiment of clubroot
Experimental subjects：Stem mustard
Prepare biological agent：Experiment biological agent is prepared according to the culture growing method of embodiment 4, is contained through plate count measure
Bacterium amount, the bacteria containing amount of bacillus amyloliquefaciens reach（1.6~3.2）×108cfu/mL；
By biological agent dilution, 20 times obtain preparation A, and by biological agent dilution, 100 times obtain preparation B, and biological agent is diluted
500 times obtain formulation C；
For medicine preparation and processing：
（1）The soil sun is heat-treated：The soil for soil of deep ploughing is poured water 30min, makes the abundant saturation of the moisture in soil, covering one
Layer plastic sheeting is sealed, and is then carried out the sun and is heat-treated 35 days, collection sun heat treatment pedotheque；
（2）Greenhouse production：Hole tray is cultivated using 72 holes, by step（1）The cave that is placed in greenhouse of sun heat treatment pedotheque
In disk, 3 ~ 4 crop in cruciferae are sowed in every cave of hole tray（Stem mustard）Vegetable seeds, take step（1）Solar heat at
Manage pedotheque covering, watering, crop in cruciferae plant（Stem mustard）For seed sprouting to 3 ~ 4 leaves are grown, it is unnecessary to cut off
Seedling, per cave retain one plant of healthy seedling, using the dilution of biological agent to crop in cruciferae plant（Stem mustard）
Seedling carries out root irrigation 1 time, and the dilution for the biological agent that each plant root adds is 10mL；
A：The dilution of biological agent is the preparation A of biological 20 times of preparation diluent；
B：The dilution of biological agent is the preparation B of biological 100 times of preparation diluent；
C：The dilution of biological agent is the formulation C of biological 500 times of preparation diluent；
D：Blank control, using clear water to crop in cruciferae plant（Stem mustard）Seedling carries out root irrigation 1 time, each plant
The clear water amount that root adds is 10mL；
Test method：In addition to dispenser sets varying level, other field management produce with conventional；Experiment sets 4 processing, 3 weights
It is multiple, Gong12Ge areas, survey production is often distinguished during harvest；
Investigation method：State of an illness overground part biological yield is investigated in stem mustard harvest time；
Experimental result
Prevention effect of the table 2. to stem mustard clubroot
The sun is heat-treated soil combination biocontrol microorganisms root irrigation, in liquid fermentation preparation（Prevent and treat crop in cruciferae clubroot
Biological agent）Bacillus amyloliquefaciensBacillus amyloliquefaciensKM 68 bacteria containing amount also can when relatively low
Good prevention effect is reached to stem mustard clubroot, works as bacillus amyloliquefaciensBacillus amyloliquefaciens
KM 68 bacteria containing amount reaches 2 × 106During cfu/mL, 76% is reached to the prevention effect of stem mustard clubroot, continues to improve biological and ecological methods to prevent plant disease, pests, and erosion
Bacteria concentration, simultaneously unobvious are improved to the prevention effect of stem mustard clubroot.
Embodiment 7：Liquid fermentation preparation（Prevent and treat the biological agent of crop in cruciferae clubroot）To crop in cruciferae
（Cabbage）The preventing and treating experiment of clubroot
Experimental subjects：Cabbage
Prepare biological agent：Experiment biological agent is prepared according to the culture growing method of embodiment 4, is contained through plate count measure
Bacterium amount, the bacteria containing amount of bacillus amyloliquefaciens reach（1.6~3.2）×108cfu/mL；
By biological agent dilution, 20 times obtain preparation A, and by biological agent dilution, 100 times obtain preparation B, and biological agent is diluted
500 times obtain formulation C；
For medicine preparation and processing：
（1）The soil sun is heat-treated：The soil for soil of deep ploughing is poured water 30min, makes the abundant saturation of the moisture in soil, covering one
Layer plastic sheeting is sealed, and is then carried out the sun and is heat-treated 35 days, collection sun heat treatment pedotheque；
（2）Greenhouse production：Hole tray is cultivated using 72 holes, by step（1）The cave that is placed in greenhouse of sun heat treatment pedotheque
In disk, 3 ~ 4 crop in cruciferae are sowed in every cave of hole tray（Cabbage）Vegetable seeds, take step（1）Solar heat
Handle pedotheque covering, watering, crop in cruciferae plant（Cabbage）Seed sprouting is cut off to 3 ~ 4 leaves are grown
Unnecessary seedling, one plant of healthy seedling is retained per cave, using the dilution of biological agent to crop in cruciferae plant（Stem mustard
Dish）Seedling carries out root irrigation 1 time, and the dilution for the biological agent that each plant root adds is 10mL；
A：The dilution of biological agent is the preparation A of biological 20 times of preparation diluent；
B：The dilution of biological agent is the preparation B of biological 100 times of preparation diluent；
C：The dilution of biological agent is the formulation C of biological 500 times of preparation diluent；
D：Blank control, using clear water to crop in cruciferae plant（Cabbage）Seedling carries out root irrigation 1 time, every plant of plant
The clear water amount that thing root adds is 10mL；
Test method：In addition to dispenser sets varying level, other field management produce with conventional；Experiment sets 4 processing, 3 weights
It is multiple, Gong12Ge areas, survey production is often distinguished during harvest；
Investigation method：State of an illness overground part biological yield is investigated in cabbage harvest time；
Experimental result
Prevention effect of the table 3. to cabbage clubroot
The sun is heat-treated soil combination biocontrol microorganisms root irrigation, in liquid fermentation preparation（Prevent and treat crop in cruciferae clubroot
Biological agent）Bacillus amyloliquefaciensBacillus amyloliquefaciensKM 68 bacteria containing amount also can when relatively low
Good prevention effect is reached to cabbage clubroot, works as bacillus amyloliquefaciensBacillus amyloliquefaciens
KM 68 bacteria containing amount reaches 2 × 106During cfu/mL, 75.36% is reached to the prevention effect of cabbage clubroot, continues to improve
Biological and ecological methods to prevent plant disease, pests, and erosion bacteria concentration, simultaneously unobvious are improved to the prevention effect of cabbage clubroot.
To sum up, sun heat treatment combines biocontrol microorganisms bacillus amyloliquefaciens has obvious preventing and treating to make to Cruciferae clubroot
With, and the yield of plant can be added, when the concentration of biocontrol microorganisms reaches 2 × 106During cfu/mL, i.e., clubroot can be reached
Good prevention effect, therefore sun processing combines bacillus amyloliquefaciensBacillus amyloliquefaciens KM 68
Preventing and treating and volume increase to Cruciferae clubroot have a good application prospect.
<110>Kunming University of Science and Technology
<120>A kind of biological agent for preventing and treating crop in cruciferae clubroot and its application
<130> 1
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1427
<212> DNA
<213> Bacillus amyloliquefaciens KM68
<400> 1
tctataatgc aagtcgagcg gacagatggg agcttgctcc ctgatgttag cggcggacgg 60
gtgagtaaca cgtgggtaac ctgcctgtaa gactgggata actccgggaa accggggcta 120
ataccggatg gttgtttgaa ccgcatggtt cagacataaa aggtggcttc ggctaccact 180
tacagatgga cccgcggcgc attagctagt tggtgaggta acggctcacc aaggcaacga 240
tgcgtagccg acctgagagg gtgatcggcc acactgggac tgagacacgg cccagactcc 300
tacgggaggc agcagtaggg aatcttccgc aatggacgaa agtctgacgg agcaacgccg 360
cgtgagtgat gaaggttttc ggatcgtaaa gctctgttgt tagggaagaa caagtgccgt 420
tcaaataggg cggcaccttg acggtaccta accagaaagc cacggctaac tacgtgccag 480
cagccgcggt aatacgtagg tggcaagcgt tgtccggaat tattgggcgt aaagggctcg 540
caggcggttt cttaagtctg atgtgaaagc ccccggctca accggggagg gtcattggaa 600
actggggaac ttgagtgcag aagaggagag tggaattcca cgtgtagcgg tgaaatgcgt 660
agagatgtgg aggaacacca gtggcgaagg cgactctctg gtctgtaact gacgctgagg 720
agcgaaagcg tggggagcga acaggattag ataccctggt agtccacgcc gtaaacgatg 780
agtgctaagt gttagggggt ttccgcccct tagtgctgca gctaacgcat taagcactcc 840
gcctggggag tacggtcgca agactgaaac tcaaaggaat tgacgggggc ccgcacaagc 900
ggtggagcat gtggtttaat tcgaagcaac gcgaagaacc ttaccaggtc ttgacatcct 960
ctgacaatcc tagagatagg acgtcccctt cgggggcaga gtgacaggtg gtgcatggtt 1020
gtcgtcagct cgtgtcgtga gatgttgggt taagtcccgc aacgagcgca acccttgatc 1080
ttagttgcca gcattcagtt gggcactcta aggtgactgc cggtgacaaa ccggaggaag 1140
gtggggatga cgtcaaatca tcatgcccct tatgacctgg gctacacacg tgctacaatg 1200
gacagaacaa agggcagcga aaccgcgagg ttaagccaat cccacaaatc tgttctcagt 1260
tcggatcgca gtctgcaact cgactgcgtg aagctggaat cgctagtaat cgcggatcag 1320
catgccgcgg tgaatacgtt cccgggcctt gtacacaccg cccgtcacac cacgagagtt 1380
tgtaacaccc gaagtcggtg aggtaacctt ttaggagcca gccgcca 1427
Claims (3)
- A kind of 1. biological agent for preventing and treating crop in cruciferae clubroot, it is characterised in that：The production bacterial strain of said preparation is Xie Dian Afnyloliquefaciens bacterial strainBacillus amyloliquefaciens KM 68, its guarantor in China typical culture collection center It is CCTCC NO to hide numbering:M 2017228, said preparation are bacillus amyloliquefaciensBacillus amyloliquefaciens The bacterium solution of the bacterial strains of KM 68.
- 2. the application process of the biological agent of the preventing and treating crop in cruciferae clubroot described in claim 1, it is characterised in that tool Body step is as follows：（1）The soil sun is heat-treated：Deep ploughing soil, pour water, seal, then carry out the sun and be heat-treated more than 30 days；（2）Greenhouse production：Acquisition step（1）The pedotheque of gained sun heat treatment is placed in the hole tray in greenhouse, in hole tray The vegetable seeds of middle sowing crop in cruciferae, the pedotheque covering for taking the sun to be heat-treated, watering, the plant of crop in cruciferae Thing seed sprouting cuts off unnecessary seedling to when growing 3 ~ 4 leaves, and one plant of healthy seedling is retained per cave, is made using biology Agent carries out root irrigation 1 time to crop in cruciferae plant seedlings.
- 3. the application process of the biological agent of preventing and treating crop in cruciferae clubroot according to claim 2, its feature exist In：Biological agent is bacillus amyloliquefaciensBacillus amyloliquefaciens The bacterium solution of the bacterial strains of KM 68, bacterium solution Gemma amount is 4 × 105~ 1×107cfu/mL。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710312344.XA CN107418910B (en) | 2017-05-05 | 2017-05-05 | Biological agent for preventing and treating clubroot of cruciferous crops and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710312344.XA CN107418910B (en) | 2017-05-05 | 2017-05-05 | Biological agent for preventing and treating clubroot of cruciferous crops and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107418910A true CN107418910A (en) | 2017-12-01 |
| CN107418910B CN107418910B (en) | 2020-08-25 |
Family
ID=60425389
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710312344.XA Active CN107418910B (en) | 2017-05-05 | 2017-05-05 | Biological agent for preventing and treating clubroot of cruciferous crops and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107418910B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109699234A (en) * | 2018-08-30 | 2019-05-03 | 四川省农业科学院植物保护研究所 | A kind of seed-coating method and capsuled seed preventing rapeseed clubroot |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101416641A (en) * | 2008-09-12 | 2009-04-29 | 云南农业大学 | Biological preparation capable of preventing and treating cruciferae club root and use thereof |
| CN102757917A (en) * | 2011-12-28 | 2012-10-31 | 浙江大学 | Biocontrol agent for preventing and curing bacterial wilt and club roots of plant as well as preparation method and application thereof |
| WO2015056813A1 (en) * | 2013-10-17 | 2015-04-23 | Ajinomoto Co., Inc. | Method for producing isoprene using bacterium |
| CN105219679A (en) * | 2015-11-02 | 2016-01-06 | 四川省农业科学院植物保护研究所 | One plant height effect Cruciferae club root biological and ecological methods to prevent plant disease, pests, and erosion bacillus amyloliquefaciens Bam22 |
| CN105230664A (en) * | 2015-11-02 | 2016-01-13 | 四川省农业科学院植物保护研究所 | Application of bacillus amyloliquefaciens Bamm22 for preventing and treating plasmodiophora brassicae |
| CN105296392A (en) * | 2015-11-02 | 2016-02-03 | 四川省农业科学院植物保护研究所 | Fermentation method of bacillus amyloliquefaciens Bam22 for preventing and treating cruciferae plasmodiophora brassicae |
| CN106399150A (en) * | 2016-07-22 | 2017-02-15 | 四川省农业科学院植物保护研究所 | Fermentation method of bacillus atrophaeus BA-7 capable of preventing and treating clubroot of cruciferae crops |
| CN106399151A (en) * | 2016-07-22 | 2017-02-15 | 四川省农业科学院植物保护研究所 | Bacillus atrophaeus BA-7 used for preventing and treating clubroot of cruciferae crops |
| CN106416835A (en) * | 2016-07-22 | 2017-02-22 | 四川省农业科学院植物保护研究所 | Method for preventing and controlling clubroot disease of cruciferous crops by means of bacillus atrophaeus BA-7 |
-
2017
- 2017-05-05 CN CN201710312344.XA patent/CN107418910B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101416641A (en) * | 2008-09-12 | 2009-04-29 | 云南农业大学 | Biological preparation capable of preventing and treating cruciferae club root and use thereof |
| CN102757917A (en) * | 2011-12-28 | 2012-10-31 | 浙江大学 | Biocontrol agent for preventing and curing bacterial wilt and club roots of plant as well as preparation method and application thereof |
| WO2015056813A1 (en) * | 2013-10-17 | 2015-04-23 | Ajinomoto Co., Inc. | Method for producing isoprene using bacterium |
| CN105219679A (en) * | 2015-11-02 | 2016-01-06 | 四川省农业科学院植物保护研究所 | One plant height effect Cruciferae club root biological and ecological methods to prevent plant disease, pests, and erosion bacillus amyloliquefaciens Bam22 |
| CN105230664A (en) * | 2015-11-02 | 2016-01-13 | 四川省农业科学院植物保护研究所 | Application of bacillus amyloliquefaciens Bamm22 for preventing and treating plasmodiophora brassicae |
| CN105296392A (en) * | 2015-11-02 | 2016-02-03 | 四川省农业科学院植物保护研究所 | Fermentation method of bacillus amyloliquefaciens Bam22 for preventing and treating cruciferae plasmodiophora brassicae |
| CN106399150A (en) * | 2016-07-22 | 2017-02-15 | 四川省农业科学院植物保护研究所 | Fermentation method of bacillus atrophaeus BA-7 capable of preventing and treating clubroot of cruciferae crops |
| CN106399151A (en) * | 2016-07-22 | 2017-02-15 | 四川省农业科学院植物保护研究所 | Bacillus atrophaeus BA-7 used for preventing and treating clubroot of cruciferae crops |
| CN106416835A (en) * | 2016-07-22 | 2017-02-22 | 四川省农业科学院植物保护研究所 | Method for preventing and controlling clubroot disease of cruciferous crops by means of bacillus atrophaeus BA-7 |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109699234A (en) * | 2018-08-30 | 2019-05-03 | 四川省农业科学院植物保护研究所 | A kind of seed-coating method and capsuled seed preventing rapeseed clubroot |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107418910B (en) | 2020-08-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105441366B (en) | Methylotrophic bacillus ZBL-1 is applied in preventing cotton verticillium wilt | |
| CN109097295A (en) | Bei Laisi Bacillus strain HN03 and its application | |
| CN105543132A (en) | Bacillus methylotrophicus YB-F7 and application thereof in preventing plant diseases | |
| CN104774106B (en) | A kind of preparation method and application of chaetomium globosum bio-bacterial manure granule | |
| CN110669691B (en) | Bacillus megaterium for preventing and treating plant nematode diseases and application thereof | |
| CN101880633A (en) | Fusarium proliferatum, and bacterium agent and application thereof | |
| CN108676761B (en) | Bacillus cereus for preventing and treating plant nematodes and application thereof | |
| CN107136122A (en) | A kind of biocontrol agent for preventing and treating the late blight of potato | |
| CN105462881A (en) | Paenibacillus polymyxa for preventing and treating vertieillium wilt in crops and application of paenibacillus polymyxa | |
| CN110218685A (en) | Biocontrol actinomycetes and application | |
| CN112342173A (en) | Bacillus belgii and application thereof | |
| CN105567600A (en) | Pathogen verticillium antagonistic bacterium and application thereof | |
| CN108641989A (en) | One plant of Methylotrophic bacillus and its application | |
| CN114196585B (en) | Burkholderia for preventing and treating tomato bacterial wilt and application thereof | |
| CN103146609B (en) | Pseudomonas fluorescens and method for preventing phytophthora capsici thereby | |
| CN108220211B (en) | Acinetobacter oleophilic NMB17 and application thereof in plant disease control | |
| CN110343621A (en) | A kind of Trichoderma asperellum strain and its application | |
| CN109439591A (en) | A kind of dead color streptomyces chromogenes SICAU417 and its application | |
| JP5374260B2 (en) | Agricultural materials | |
| CN111778174B (en) | Bacillus subtilis with inhibiting effect on citrus sand skin disease and screening method thereof | |
| CN107043719B (en) | bacillus amyloliquefaciens HMB28353 and application thereof | |
| CN108753666A (en) | The preparation and application of one plant of salt tolerant Antagonistic Fungi M4-1 and its microbial inoculum | |
| CN108913625A (en) | Salt tolerant streptomycete, its microbial inoculum and its microbial inoculum are promoting the application in plant growth | |
| CN116694505A (en) | Streptomyces crimson capable of preventing and treating clubroot and application thereof | |
| CN107418910A (en) | A kind of biological agent for preventing and treating crop in cruciferae clubroot and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |